Download PDF
Thromb Haemost 1996; 75(03): 491-496
DOI: 10.1055/s-0038-1650303
Original Article
Schattauer GmbH Stuttgart

Physico-Chemical Properties and the Structure of Dermatan Sulfate Fractions Purified from Plasma after Oral Administration in Healthy Human Volunteers

Nicola Volpi
The Department of “Biologia Animale”, Chair of Biological Chemistry, University of Modena, Italy
› Author Affiliations
Further Information

Publication History

Received 26 September 1995

Accepted after revision 20 November 1995

Publication Date:
26 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Dermatan sulfate (DS) was administered by oral route in healthy human volunteers. The structure, physico-chemical properties and biological activity of DS purified from human plasma after oral administration were studied and compared with those of native DS.

DS extracted and purified from pig mucosa has a relative molecular mass (Mr) of about 23,100 and is composed of about 10% nonsulfated disaccharide, 80% monosulfated disaccharides and about 10% disulfat-ed disaccharides, with a sulfate to carboxyl ratio of 1.00 and a heparin cofactor II (HCII) activity of about 160 units/mg. This native polysaccharide is composed of about 94% iduronic acid.

One gram of native DS was orally administered to five healthy human volunteers, and 50 ml of blood were collected after 4 h. DS possibly present in plasma after oral administration was extracted and purified. About 130 ± 42 Μg of DS per 50 ml of blood were detected by agarose-gel electrophoresis and DMB assay. This DS shows a broad Mr range. After oral absorption, substantial amounts of species with a Mr of about 7,500 are detected in blood but chains with Mr ranging from 7,500 to 20,000 are also found. Moreover, some very low-Mr species are detected, with a prevalence of disaccharides. After oral absorption, DS is sulfated above all in position 4 of the N-acetyl-galactosamine (60%), with a sulfate to carboxyl ratio of about 0.64, demonstrating that DS is desulfated during or after oral absorption by about 30-40%. A small amount of disulfated disaccharide (in particular 2,4-disulfated, 1.4%) is preserved from catabolic processes, as DS extracted from human plasma is able to inhibit thrombin activity mediated by HCII (about 16 U/mg).

 

  • References

  • 1 Mammen EF, Walenga JM, Fareed J. Seminars in thrombosis and hemostasis. Thieme Medical Publishers; New York, Stuttgard: 1991. 17 1-2
  • 2 Scott JE. Dermatan sulphate proteoglycans. Portland Press; London, U.K.: 1993
  • 3 Al-Hakim A, Linhardt RJ. Capillary electrophoresis for the analysis of chondroitin sulfate- and dermatan sulfate-derived disaccharides. Analyt Biochem 1991; 195: 68-73
    CrossrefPubMedGoogle Scholar
  • 4 Maimone MM, Tollefsen DM. Structure of a dermatan sulfate hexasaccha-ride that binds to heparin cofactor II with high affinity. J Biol Chem 1990; 265: 18263-18271
    PubMedGoogle Scholar
  • 5 Volpi N. Dermatan sulfate from beef mucosa: structure, physicochemical and biological properties of fractions prepared by chemical depolymerization and anion-exchange chromatography. Carb Res 1994; 255: 133-144
    CrossrefPubMedGoogle Scholar
  • 6 Ryan KE, Lane DA, Flynn A, Ireland H, Boisclair M, Shepperd J, Curtis JR. Antithrombotic properties of dermatan sulphate (MF 701) in haemodialysis for chronic renal failure. Thromb Haemost 1992; 68: 563-569
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Barbanti M, Calanni F, Milani MR, Marchi E, Semeraro N, Colucci M. Therapeutic effect of a low molecular weight dermatan sulphate (desmin 370) in rat venous thrombosis - Evidence for an anticoagulant-independent mechanism. Thromb Haemost 1993; 69: 147-151
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Bendayan P, Boccalon H, Dupouy D, Boneu B. Dermatan sulfate is a more potent inhibitor of clot-bound thrombin than unfractionated and low molecular weight heparins. Thromb Haemost 1994; 71: 576-580
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Fernandez F, Van Ryn J, Ofosu FA, Hirsh J, Buchanan MR. The haemor-rhagic and antithrombotic effects of dermatan sulphate. Br J Haematol 1986; 64: 309-317
    CrossrefPubMedGoogle Scholar
  • 10 Dol F, Houin G, Rostin M, Montastruc JL, Dupouy D, Gianese F, Sié P, Boneu B. Pharmacodynamics and pharmacokinetics of dermatan sulfate in humans. Blood 1989; 74: 1577-1582
    PubMedGoogle Scholar
  • 11 Dawes J, McLaren M, Forbes C, Belch JJF, Lane DA, Bray B, McEwen J, Houin G, Gianese F. The pharmacokinetics of dermatan sulphate MF701 in healthy human volunteers. Br J Clin Pharmac 1991; 32: 361-366
    CrossrefPubMedGoogle Scholar
  • 12 Dawes J, Hodson BA, Pepper DS. The absorption, clearance and metabolic fate of dermatan sulphate administered to man - studies using a radioio-dinated derivative. Thromb Haemost 1989; 62: 945-949
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Imbimbo BP, Sié P, Agnelli G, Saivin S, Dupouy D, Damiani M, Houin G, Gianese F. Intramuscular dermatan sulfate MF701 in patients with hip fracture: relationship between pharmacokinetics and antithrombotic efficacy. Thromb Haemost 1994; 71: 553-557
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Volpi N. Fractionation of heparin, dermatan sulfate, and chondroitin sulfate by sequential precipitation: a method to purify a single glycosaminoglycan species from a mixture. Analyt Biochem 1994; 218: 382-391
    CrossrefPubMedGoogle Scholar
  • 15 Shively JE, Conrad HE. Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry 1976; 15: 3932-3942
    CrossrefPubMedGoogle Scholar
  • 16 Poblacion CA, Michelacci YM. Structural differences of dermatan sulfates from different origins. Carb Res 1986; 147: 87-100
    CrossrefPubMedGoogle Scholar
  • 17 Volpi N. “Fast moving” and “slow moving” heparins, dermatan sulfate, and chondroitin sulfate: qualitative and quantitative analysis by agarose-gel electrophoresis. Carb Res 1993; 247: 263-278
    CrossrefPubMedGoogle Scholar
  • 18 Cappelletti R, De Rosso M, Chiarugu VP. A new electrophoretic method for the complete separation of all known animal glycosaminoglycans in a monodimensional run. Analyt Biochem 1979; 99: 311-315
    CrossrefPubMedGoogle Scholar
  • 19 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-phenol method. J Biol Chem 1951; 193: 317-323
    PubMedGoogle Scholar
  • 20 Volpi N, Cusmano M, Venturelli T. Qualitative and quantitative studies of heparin and chondroitin sulfates in normal human plasma. Biochim Biophys Acta 1995; 1243: 49-58
    CrossrefPubMedGoogle Scholar
  • 21 Al-Hakim A, Linhardt RJ. Electrophoresis and detection of nanogram quantities of exogenous and endogenous glycosaminoglycans in biological fluids. Appl and Theor Electrophoresis 1991; 1: 305-312
    PubMedGoogle Scholar
  • 22 Staprans I, Felts JM. Isolation and characterization of glycosaminoglycans in human plasma. J Clin Invest 1985; 76: 1984-1991
    CrossrefPubMedGoogle Scholar
  • 23 Rice KG, Rottink MK, Linhardt RJ. Fractionation of heparin-derived oligosaccharides by gradient poly aery lamide-gel electrophoresis. Biochem J 1987; 244: 515-522
    CrossrefPubMedGoogle Scholar
  • 24 Heinegard D, Sommarin Y. Isolation and characterization of proteoglycans. Methods in Enzymol 1987; 144: 319-372
    PubMedGoogle Scholar
  • 25 Whitley CB, Ridnour MD, Draper KA, Dutton CM, Neglia JP. Diagnostic test for mucopolysaccharidosis. I. Direct method for quantifying excessive urinary glycosaminoglycan excretion. Clin Chem 1989; 35: 374-379
    PubMedGoogle Scholar
  • 26 Dupouy D, Sié P, Dol F, Boneu B. A simple method to measure dermatan sulfate at submicrogram concentrations in plasma. Thromb Haemost 1988; 60: 236-239
    Article in Thieme ConnectPubMedGoogle Scholar
  • 27 Volpi N. Characterization of heparins with different relative molecular masses (from 11600 to 1600) by various analytical techniques. J Chrom Biomed Appl 1993; 622: 13-20
    CrossrefPubMedGoogle Scholar
  • 28 Pescador R, Diamantini G, Mantovani M, Malandrino S, Riva A, Casu B, Oreste P. Absorption by the rat intestinal tract of fluorescein-labelled pig duodenal glycosaminoglycans. Arzneim-Forsch 1980; 30: 1893-1896
    PubMedGoogle Scholar
  • 29 Larsen AK, Lund DP, Langer R, Folkman J. Oral heparin results in the appearance of heparin fragments in the plasma of rats. Proc Natl Acad Sci USA 1986; 83: 2964-2968
    CrossrefPubMedGoogle Scholar
  • 30 Palmieri L, Conte A, Giovannini L, Lualdi P, Ronca G. Metabolic fate of exogenous chondroitin sulfate in the experimental animal. Arzneim-Forsch 1990; 40: 319-323
    PubMedGoogle Scholar
  • 31 Conte A, de Bemardi M, Palmierig L, Lualdi P, Mautone G, Ronca G. Metabolic fate of exogenous chondroitin sulfate in man. Arzneim-Forsch 1991; 41: 768-772
    PubMedGoogle Scholar
  • 32 Baici A, Horler D, Moser B, Hofer HO, Fehr K, Wagenhauser FJ. Analysis of glycosaminoglycans in human serum after oral administration of chondroitin sulfate. Rheumatol Int 1992; 12: 81-88
    CrossrefPubMedGoogle Scholar
  • 33 Crepaldi G, Rossi A, Coscetti G, Abbruzzese E, Calveri U, Calabrò A. Sulodexide oral administration influences blood viscosity and fibrinolysis. Drugs Exptl Clin Res 1992; 18: 189-195
    PubMedGoogle Scholar
  • 34 Scagnol I, Fumagalli G, Andriuoli G. Anticoagulant and antithrombotic activity of heparin salts by intraduodenal route in rabbits. Thromb Res 1992; 68: 195-200
    CrossrefPubMedGoogle Scholar
  • 35 Jaques LB, Hiebert LM, Wice SM. Evidence from endothelium of gastric absorption of heparin and of dextran sulfates 8000. J Lab Clin 1991; 117: 122-130
    PubMedGoogle Scholar
  • 36 Fioravanti A, Franci A, Anselmi F, Fattorini L, Marcolongo R. Clinical efficacy and tolerance of galactosaminoglucuronoglycan sulfate in the treatment of osteoarthritis. Drugs Exptl Clin Res 1991; XVII: 41-44
    PubMedGoogle Scholar
  • 37 Vasdev S, Sampson CA, Longerich L, Parai S. Oral heparin prevents hypertension and elevated cytosolic calcium in salt-sensitive rats. Artery 1992; 19: 225-245
    PubMedGoogle Scholar
  • 38 Baggio B, Gambaro G, Marzaro G, Marchini F, Borsatti A, Crepaldi G. Effects of the oral administration of glycosaminoglycans on cellular abnormalities associated with idiopathic calcium oxalate nephrolithiasis. Eur J Clin Pharm 1991; 40: 237-240
    CrossrefPubMedGoogle Scholar